Cell surface glycans are critical for interactions that initiate many enteric infections. This new grant application proposes translational studies to test the hypothesis that the complex interactions between glycan expression, neonatal development and human milk oligosaccharides affect neonatal enteric infections. Characterization of a rotavirus strain that almost exclusively infects neonates in India and new data that human rotaviruses can initiate infection by binding to histo blood group antigens (HBGAs) provide the foundation for this study. We propose: (1) to characterize intestinal glycans that are novel binding partners in rotavirus infectivity by analyzing the interactions between a rotavirus strain that infects neonates and glycans with the Gal?1-4GlcNAc motif including H type II HBGA, and (2) to determine the role of bioactive components of breast milk such as human milk oligosaccharides (HMO) and functional antibodies that competitively bind to the virus and block viral binding to intestinal glycans in protection from neonatal rotavirus infection and disease. Fo the first goal, we will identify critical glycan partners that bind to the neonatal rotavirus strai and are required for rotavirus infectivity. The second goal will be achieved by studying samples to be collected from neonates and their mothers in a nursery where incidence of infection is reproducibly high (nearly 50%). We predict that (a) structural analogues to intestinal glycans in breast milk play a protective role in neonatal infections by competitively inhibiting the binding o the pathogen to these glycans or (b) functional antibodies in breast milk inhibit the binding of th virus to glycans. A multidisciplinary team with expertise in epidemiology and field studies of enteric infection in children including neonates, molecular virology and viral pathogenesis, and in glycobiology and human milk oligosaccharides, will apply technological advances in glycobiology and human intestinal organoids to obtain new fundamental data on glycan expression in neonates and on whether human milk bioactive components specifically block neonatal viral infection. The information from this study may discover the first correlate of protection for rotavirus and will provide mechanistic insight into glycan expression during intestinal development of the neonate. This is likely to be significant and relevant to other neonatal infections or disease conditions where glycans play a significant role.
Neonatal enteric infections are important globally and accumulating evidence supports the idea that the expression of glycan receptors on susceptible intestinal epithelial cells as well as oligosaccharides in mother's milk may be critical in determining if infection occurs and if the outcome of such infection is asymptomatic or symptomatic. Proving this hypothesis can be challenging when the incidence of such infections is low and if multiple strains of a microbe cause such infections. This project will test this hypothesis by studying a unique rotavirus strain that is already characterized to bind to at least one glycan, whose expression increases virus infectivity in vitro and whose core structure is part of human milk oligosaccharides. This study is being carried out in a setting in India where the pathogen routinely infects neonates at a high incidence (nearly 50%), thus allowing collection of sufficient relevant samples from a large cohort of neonates and their mothers to determine whether, how and what specific human milk oligosaccharides protect the breast-fed neonate from infection or illness. These studies will define the role of different glycans for virus infectvity in cultured cells including human intestinal organoids and determine if critical interactions between virus, host cells and breast milk that impact the health of human neonates can be molecularly understood.
|Hu, Liya; Ramani, Sasirekha; Czako, Rita et al. (2015) Structural basis of glycan specificity in neonate-specific bovine-human reassortant rotavirus. Nat Commun 6:8346|
|Yu, Ying; Lasanajak, Yi; Song, Xuezheng et al. (2014) Human milk contains novel glycans that are potential decoy receptors for neonatal rotaviruses. Mol Cell Proteomics 13:2944-60|
|Ashline, David J; Yu, Ying; Lasanajak, Yi et al. (2014) Structural characterization by multistage mass spectrometry (MSn) of human milk glycans recognized by human rotaviruses. Mol Cell Proteomics 13:2961-74|
|Venkataram Prasad, B V; Shanker, Sreejesh; Hu, Liya et al. (2014) Structural basis of glycan interaction in gastroenteric viral pathogens. Curr Opin Virol 7:119-27|
|Ramani, Sasirekha; Cortes-Penfield, Nicolas W; Hu, Liya et al. (2013) The VP8* domain of neonatal rotavirus strain G10P binds to type II precursor glycans. J Virol 87:7255-64|